İskemi/reperfüzyon hasarında Apelin'in etkilerinin araştırılması
Küçük Resim Yok
Tarih
2021
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi, Tıp Fakültesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Plastik ve Rekonstrüktif Cerrahi uygulamasında travma, tümör rezeksiyonu vb. durumlarda doku kaybı sonrası flep cerrahisi çok sık olarak uygulanmaktadır. Zamanla teknoloji ilerlemesiyle kullanılan aletlerde geliştirilmiş ve zamanla flep cerrahisinde çığır açan değişiklikler olmuştur. Mikroskopun bulunması ve cerrahide kullanılması mikrocerrahi tanımını ortaya çıkarmıştır. Alexis Carrel tarafından tanımlanan end-to-end anastomoz için
kullanılan triangulasyon metodu gibi bir çok damar onarma şekli tanımlanmaya başlamıştır (1). Böylece flep cerrahisi de gelişmiş, doku defektlerinin kapatılmasındaki başarı da buna paralel olarak ilerlemiştir. Uzun zamandır plastik cerrahların ilgi odağında olan doku defektlerine flep uygulaması esnasında teknik ve aletlerdeki ilerlemelere rağmen zorlu süreçlerin başında iskemi reperfüzyon hasarı gelmektedir. İskemi reperfüzyon hasarının etkilerinden kurtulmak için çeşitli madde enjeksiyonları tanımlanmış, cerrahi teknikte farklı yaklaşımlar denenmiştir ancak bu konu flep cerrahisinin zorlayıcı yanlarından biri olmaya devam etmektedir (2). Bu çalışmamız, sıçan modellerinde serbest inguinal fleplerde oluşan iskemi reperfüzyon hasarı tablosunun jel formülasyonunda hazırlanan apelin’in azaltıcı etkisi olup olmadığı araştırılmıştır. Çalışmamızdaki amaç, daha önce hayvan deneylerinde kalp, beyin, deride iskemik hasara karşı etkili olduğu gösterilen apelinin jel formülasyonuyla uzun süreli dokuya salınımın flep içinde ve flep etrafı dokuda oluşan anormal değişiklikleri azaltabileceği, hasarı ne boyutta engelleyeceği, damar oluşumuna etkisiyle dokunun iskemiye karşı yanıtının ne yönde etkileyeceğinin tespit edilmesidir. Deney modelimizde biyoistatistiksel olarak anlamlı sonuç almak için 28 adet Wistar albino türü sıçan kullanıldı. 4 ayrı grup oluşturuldu. Anestezi için önce indüksiyon odasında gaz inhalasyonu inhaler isofluran 3 lt/dk dan ve 5 lt/dk dan oksijen gazı uygulandı. Sıçanlara sonrasında ketamin ve ksilazin ile anestezi uygulandı. Devamında 2 lt/dk dan isofluran ve 4 lt/dk dan oksijen gazı ile idame genel anestezi sağlanmış oldu. Anestezi sonrası deneyin yapılacağı inguinal bölgede %70 etanol ve ardından steril serum fizyolojik ile temizlik işlemi yapıldı. Birinci grupta herhangi bir iskemik girişim yapılmayarak inguinal flep kaldırıldı. Daha sonra flep flow-through olacak şekilde femoral arter ve ven üzerinden kaldırıldı ve herhangi bir iskemik işlem uygulanmadı. İkinci gruptaki sıçanlarda ise aynı şekilde inguinal flep kaldırıldı. 2-saat iskemi süresi sonrası damar anastomozu gerçekleştirildi. Üçüncü gruptaki sıçanlarda, aynı şekilde serbest flep inguinal bölgeden kaldırıldı, 2-saat süreyle iskemi uygulandı ve damar
anastomozu sonrası subkutan planda apelin içermeyen jel formülayonu uygulandı. Dördüncü grupta iste aynı işlemler apelin içeren jel formülasyonu ile beraber uygulandı. Yedi gün sonra tüm flepler kontrol edilerek tamamen alındı ve makro düzeyde incelemeler sonrası formaldehit solüsyonu içerisine konularak fikse edildi ve rutin histolojik takip prosedürü uygulandı. Histolojik değerlendirmede hemotoksilen-eosin ve Mallory azan ile boyanmış örnekler histopatolojik olarak Modifiye Verhofstad Skorlaması ile değerlendirildi. Ayrıca anti e-NOS ve anti-VEGF immunohistokimyasal boyamaları gerçekleştirildi. Örneklere modifiye verhostad skorlaması yapılarak skorların ortalamaları hesaplandığında 4.grubun 2.gruba göre daha az iskemi-reperfüzyon skoru aldığı görüldü. Bu bulgularla iskemi-reperfüzyon hasarı gelişmesini önlemek amacıyla apelinin jel formülasyonunun kullanımının anlamlı etkisinin olduğu kanaatine varıldı.
In Plastic and Reconstructive Surgery practice, flap surgery is performed very frequently after tissue loss in cases such as trauma, tumor resection, etc. Over time, the tools used with the advancement of technology were also developed and over time, there were groundbreaking changes in flap surgery. The presence of the microscope and its use in surgery established the definition of microsurgery. Many forms of vascular repair have been identified, such as the trianguation method used for end-to-end anastomosis described by Alexis Carrel (1). Thus, the success of flap surgery in closing advanced tissue defects has evolved in parallel with this. Ischemia reperfusion damage is one of the most challenging processes despite advances in techniques and instruments during the application of flaps to tissue defects that have long been the focus of attention of plastic surgeons. Various injections of substances have been defined to overcome the effects of ischemia reperfusion injury, different approaches have been tried in surgical technique, but this issue remains one of the challenging aspects of flap surgery(2). In this study, it was investigated whether apelin prepared in gel formulation had a mitigating effect on the ischemia reperfusion damage in free inguinal flaps in a rat model. The aim of our study is to determine The aim of our study is that long-term release to the tissue with the gel formulation of apelin, which has been shown to be effective against ischemic damage to the heart, brain and skin in animal experiments, can reduce the abnormal changes occurring in the flap and the tissue around the flap, to what extent it will prevent the damage, and in what direction the tissue's response will be to ischemia with its effect on vessel formation, determining its impact. In our experimental model, 28 Wistar albino type rats were used to obtain biostatistically significant results. 4 separate groups were created. For anesthesia, inhaler isofluran and oxygen gas mix between 3 lt/min and 5 lt/min was applied in the induction chamber. Rats were then anesthetised with ketaminele xylazin. Afterwards, general anesthesia was provided with 2 lt/min isofluran and 4 lt/min oxygen. After anesthesia induction, inguinal area was cleaned with 70% ethanol and sterile serum phsyiologique. In the first group, the inguinal flap was elevated without any ischemic insult. The flap was then elevated till femoral artery and vein so that it would be a flow-through flap without any ischemia. In rats in the second group, the inguinal flap was elevated the same way. Vascular anastomoses were performed after a 2 hours of ischemia period. In rats of the third group, the free flap was elevated from the inguinal region, ischemia was applied for 2-hours, and Apelin-free gel formulation was applied to the subcutanous plane after vascular anastomosis. In the fourth group, the same procedures were done together with the gel formulation containing apelin. After seven days, all the flaps were checked, and excised, and after macroscopic examinations, flaps were put into formaldehyde solution and routine histological follow-up. In histological evaluation, samples stained with hemotoxylene-eosin and Mallory azan were evaluated histopathologically with modified Verhofstad Scoring. In addition, sections were stained with anti e-NOS and anti-VEGF immunohistochemical dyes. When the averages of the scores were calculated by performing modified verhostad scoring on the samples, it was seen that the 4th group received fewer ischemia-reperfusion scores than the 2nd group. With these findings, it was concluded that the use of apelin gel formulation had a meaningful effect in order to prevent the development of ischemia-reperfusion damage.
In Plastic and Reconstructive Surgery practice, flap surgery is performed very frequently after tissue loss in cases such as trauma, tumor resection, etc. Over time, the tools used with the advancement of technology were also developed and over time, there were groundbreaking changes in flap surgery. The presence of the microscope and its use in surgery established the definition of microsurgery. Many forms of vascular repair have been identified, such as the trianguation method used for end-to-end anastomosis described by Alexis Carrel (1). Thus, the success of flap surgery in closing advanced tissue defects has evolved in parallel with this. Ischemia reperfusion damage is one of the most challenging processes despite advances in techniques and instruments during the application of flaps to tissue defects that have long been the focus of attention of plastic surgeons. Various injections of substances have been defined to overcome the effects of ischemia reperfusion injury, different approaches have been tried in surgical technique, but this issue remains one of the challenging aspects of flap surgery(2). In this study, it was investigated whether apelin prepared in gel formulation had a mitigating effect on the ischemia reperfusion damage in free inguinal flaps in a rat model. The aim of our study is to determine The aim of our study is that long-term release to the tissue with the gel formulation of apelin, which has been shown to be effective against ischemic damage to the heart, brain and skin in animal experiments, can reduce the abnormal changes occurring in the flap and the tissue around the flap, to what extent it will prevent the damage, and in what direction the tissue's response will be to ischemia with its effect on vessel formation, determining its impact. In our experimental model, 28 Wistar albino type rats were used to obtain biostatistically significant results. 4 separate groups were created. For anesthesia, inhaler isofluran and oxygen gas mix between 3 lt/min and 5 lt/min was applied in the induction chamber. Rats were then anesthetised with ketaminele xylazin. Afterwards, general anesthesia was provided with 2 lt/min isofluran and 4 lt/min oxygen. After anesthesia induction, inguinal area was cleaned with 70% ethanol and sterile serum phsyiologique. In the first group, the inguinal flap was elevated without any ischemic insult. The flap was then elevated till femoral artery and vein so that it would be a flow-through flap without any ischemia. In rats in the second group, the inguinal flap was elevated the same way. Vascular anastomoses were performed after a 2 hours of ischemia period. In rats of the third group, the free flap was elevated from the inguinal region, ischemia was applied for 2-hours, and Apelin-free gel formulation was applied to the subcutanous plane after vascular anastomosis. In the fourth group, the same procedures were done together with the gel formulation containing apelin. After seven days, all the flaps were checked, and excised, and after macroscopic examinations, flaps were put into formaldehyde solution and routine histological follow-up. In histological evaluation, samples stained with hemotoxylene-eosin and Mallory azan were evaluated histopathologically with modified Verhofstad Scoring. In addition, sections were stained with anti e-NOS and anti-VEGF immunohistochemical dyes. When the averages of the scores were calculated by performing modified verhostad scoring on the samples, it was seen that the 4th group received fewer ischemia-reperfusion scores than the 2nd group. With these findings, it was concluded that the use of apelin gel formulation had a meaningful effect in order to prevent the development of ischemia-reperfusion damage.
Açıklama
Anahtar Kelimeler
Apelin, İskemi-Reperfüzyon Hasarı, Serbest Flep, In Situ Hidrojel, Apelin, Free Flap, Ischemia-Reperfusion Injury, In Situ Hydrogel
